IL-12 inhibits TNF-α induced osteoclastogenesis via a T cell-independent mechanism in vivo

It has been reported that TNF-α plays an important role in bone resorption in pathological conditions. IL-12, which is a T cell mediator, is also an important inflammatory cytokine. We previously reported that IL-12 induces apoptosis in bone marrow cells treated with TNF-α in vitro via an interaction between TNF-α-induced Fas and IL-12-induced Fas ligand (FasL), and that, as a result, osteoclastogenesis is inhibited. The purpose of this study was to investigate the effects of IL-12 on TNF-α-mediated osteoclastogenesis in vivo. We administered TNF-α with and without IL-12 into the supracalvaria in mice. The numbers of osteoclasts in the sutures in the calvaria were higher in mice administered TNF-α than in control mice not administered TNF-α. The numbers of osteoclasts in mice administered both TNF-α and IL-12 were lower than those in mice administered only TNF-α. Next, we determined the levels of mRNAs for cathepsin K and tartrate-resistant acid phosphatase (TRAP). mRNA levels were increased in mice administered TNF-α compared with control mice, but not in mice administered both TNF-α and IL-12. We also evaluated the amounts of tartrate-resistant acid phosphatase 5b (TRACP 5b) in mouse sera. The levels of TRACP 5b in mice administered TNF-α were higher than those in control mice. On the other hand, in mice administered both TNF-α and IL-12, the levels were lower than those in mice administered TNF-α alone. Fas and FasL expression levels were analyzed by real-time RT-PCR. The levels of Fas mRNA were increased in the calvaria of mice administered TNF-α compared with control mice, while those of FasL mRNAs were increased in the calvaria of mice administered IL-12. In TdT-mediated dUTP-biotin nick end-labeling (TUNEL) assays, many apoptotic cells were found in the sutures in the calvaria of mice administered both TNF-α and IL-12. IL-12 also inhibited TNF-α-induced osteoclastogenesis in mice whose T cells were blocked by anti-CD4 and anti-CD8 antibodies. These results suggest that IL-12 inhibits TNF-α-mediated osteoclastogenesis and induces apoptotic changes through an interaction between TNF-α-induced Fas and IL-12-induced FasL, in vivo, via a T cell-independent mechanism.     

前列腺癌和良性前列腺增生组织中M3受体表达及意义

目的 探讨前列腺组织中胆碱能毒蕈碱受体M3亚型表达在前列腺肿瘤发生发展中的作用.方法 收集正常前列腺、良性前列腺增生、前列腺癌组织标本各36例.应用RT-PCR方法分别测定标本组织中M3受体、血管内皮生长因子(VEGF)的表达情况;蛋白质印迹与免疫组织化学染色方法测定各组标本中M3、VEGF、人白细胞分化抗原34(CD34)的蛋白表达.图像分析系统检测数据,以相对表达强度(目的 物质表达强度/β-actin表达强度)作为统计学分析参数.数据统计分析 计数资料采用方差分析,分类变量资料采用χ2检验,相关性分析采用Pearson方法.结果 前列腺癌组织标本M3受体基因相对表达强度0.8354±0.1897、VEGF基因表达0.7824±0.2047,明显高于良性前列腺增生组织(0.6735±0.1603、0.6021±0.1637)和正常前列腺组织(0.5425±0.1629、0.3436±0.1581),组间差异有统计学意义(P<0.001),3种组织中M3受体与VEGF基因表达呈直线正相关关系(r=0.4999,P<0.001).前列腺癌组织标本M3受体蛋白表达0.4777±0.1638、VEGF蛋白表达0.5718±0.2245,明显高于良性前列腺增生组织(0.3655±0.1474、0.4342±0.1538)和正常前列腺组织(0.2659±0.1076、0.3380±0.1527),组间差异有统计学意义(P<0.05).结论 M3受体表达与前列腺肿瘤的发生发展有关.     

PI3K-Akt-eNOS信号转导通路在吗啡促人脐静脉内皮细胞一氧化氮合成中的作用

目的 评价磷脂酰肌醇-3激酶-蛋白质丝氨酸苏氨酸激酶-内皮型一氧化氮合酶(PI3K-Akt-eNOS)信号转导通路在吗啡对促人脐静脉内皮细胞一氧化氮(NO)合成中的作用.方法 体外培养人脐静脉内皮细胞,实验Ⅰ:人脐静脉内皮细胞随机分为4组,每组6孔:C组和M1～3组,C组不予任何处理;M1～3组加入吗啡,终浓度为1 μmol/L,分别孵育3、6和12 h,随后测定NO含量;实验Ⅱ:人脐静脉内皮细胞随机分为4组,每组6孔:C组和M1～3组,C组不予任何处理;M1～3组加入吗啡,终浓度分别为0.01、1和100 μmol/L,孵育6 h,随后测定NO含量;实验Ⅲ:人脐静脉内皮细胞随机分为5组,每组6孔:C组、M组、MW组、MS组和ML组,C组不予任何处理;M组加入吗啡,终浓度1 μmol/L;MW组先加入wortmannin(PI3K特异性抑制剂),15 min后加入吗啡,终浓度分别为5、1μmol/L;MS组先加入SH-5(Akt特异性抑制剂),15 rain后加入吗啡,终浓度分别为10、1 μmol/L;ML组先加入L-NAME(eNOS特异性抑制剂),15 min后加入吗啡,终浓度分别为0.5、1 μmol/L,各组孵育6 h后,测定NO含量.结果 实验Ⅰ结果 :随吗啡孵育时间延长,促脐静脉内皮细胞NO合成的作用增强(P<0.05);实验Ⅱ结果 :随吗啡浓度升高,促脐静脉内皮细胞NO合成的作用增强(P<0.05);实验Ⅲ结果 :与C组比较,M组人脐静脉内皮细胞NO含量升高(P<0.05),MW组、MS组和ML组差异无统计学意义(P>0.05);与M组比较,MW组、MS组和ML组人脐静脉内皮细胞NO含量降低(P<0.05).结论 吗啡促人脐静脉内皮细胞NO合成呈浓度和时间依赖性,其机制可能与激活PI3K-Akt-eNOS信号转导通路有关.     

The Role of Land Use Patterns in Limiting the Spread of Equine Influenza in Queensland During the 2007 Epidemic

In 2007, an epizootic of equine influenza (EI) occurred in Australia, involving parts of the states of Queensland and New South Wales. Following an extensive control program, the disease was eradicated within 4 months, after infecting more than 75 000 horses on over 10 000 properties. In Queensland, examination of land use patterns revealed that the majority of infected premises (89.5%) were located in one of three land use classes viz. rural residential, residential‐unspecified or grazing natural vegetation. All nine clusters of infection in Queensland were surrounded to some degree by parcels of land whose designated use precluded the presence of horses. In four clusters, these horse‐free areas completely enclosed the cluster and spread was limited to a relatively small area whereas the remaining five clusters were not completely surrounded by horse‐free buffers and spread was much more extensive. The cumulative incidence of infection with EI dropped from 41% of properties in rural‐residential areas to 4% of properties located further than 5 km from rural‐residential areas. This decrease was associated with a 10‐fold increase in the median size of properties and a 5‐fold decrease in the density of horses on those premises. There was a strong negative correlation between the cumulative incidence of infection and the median property size (R = −0.91, P < 0.05) and a positive correlation with the horse density on those properties (R = 0.93, P < 0.01).     

Validation of a new method for determination of cup orientation in THA

Our goal was to validate accuracy, consistency, and reproducibility/reliability of a new method for determining cup orientation in total hip arthroplasty (THA). This method allows matching the 3D‐model from CT images or slices with the projected pelvis on an anteroposterior pelvic radiograph using a fully automated registration procedure. Cup orientation (inclination and anteversion) is calculated relative to the anterior pelvic plane, corrected for individual malposition of the pelvis during radiograph acquisition. Measurements on blinded and randomized radiographs of 80 cadaver and 327 patient hips were investigated. The method showed a mean accuracy of 0.7 ± 1.7° (?3.7° to 4.0°) for inclination and 1.2 ± 2.4° (?5.3° to 5.6°) for anteversion in the cadaver trials and 1.7 ± 1.7° (?4.6° to 5.5°) for inclination and 0.9 ± 2.8° (?5.2° to 5.7°) for anteversion in the clinical data when compared to CT‐based measurements. No systematic errors in accuracy were detected with the Bland–Altman analysis. The software consistency and the reproducibility/reliability were very good. This software is an accurate, consistent, reliable, and reproducible method to measure cup orientation in THA using a sophisticated 2D/3D‐matching technique. Its robust and accurate matching algorithm can be expanded to statistical models. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1583–1588, 2009     

T2-Weighted Liver MRI Using the MultiVane Technique at 3T: Comparison with Conventional T2-Weighted MRI

Objective

To assess the value of applying MultiVane to liver T2-weighted imaging (T2WI) compared with conventional T2WIs with emphasis on detection of focal liver lesions.

Materials and Methods

Seventy-eight patients (43 men and 35 women) with 86 hepatic lesions and 20 pancreatico-biliary diseases underwent MRI including T2WIs acquired using breath-hold (BH), respiratory-triggered (RT), and MultiVane technique at 3T. Two reviewers evaluated each T2WI with respect to artefacts, organ sharpness, and conspicuity of intrahepatic vessels, hilar duct, and main lesion using five-point scales, and made pairwise comparisons between T2WI sequences for these categories. Diagnostic accuracy (Az) and sensitivity for hepatic lesion detection were evaluated using alternative free-response receiver operating characteristic analysis.

Results

MultiVane T2WI was significantly better than BH-T2WI or RT-T2WI for organ sharpness and conspicuity of intrahepatic vessels and main lesion in both separate reviews and pairwise comparisons (p < 0.001). With regard to motion artefacts, MultiVane T2WI or BH-T2WI was better than RT-T2WI (p < 0.001). Conspicuity of hilar duct was better with BH-T2WI than with MultiVane T2WI (p = 0.030) or RT-T2WI (p < 0.001). For detection of 86 hepatic lesions, sensitivity (mean, 97.7%) of MultiVane T2WI was significantly higher than that of BH-T2WI (mean, 89.5%) (p = 0.008) or RT-T2WI (mean, 84.9%) (p = 0.001).

Conclusion

Applying the MultiVane technique to T2WI of the liver is a promising approach to improving image quality that results in increased detection of focal liver lesions compared with conventional T2WI.     

Sirt1在急性炎症状态下糖尿病小鼠肾损伤中的作用

目的 探究Sirt1在急性炎症状态下糖尿病小鼠肾损伤中的作用及分子机制。方法选择SPF级C57BL/6J雄性小鼠40只,8周龄,体重20～25 g。采用随机数字表法将小鼠分为五组：对照组(C组)、糖尿病组(D组)、脂多糖(LPS)+糖尿病组(L组)、LPS+糖尿病+Sirt1阻断剂EX527组(E组)和LPS+糖尿病+Sirt1激动剂银杏黄酮苷元组(G组),每组8只。糖尿病小鼠模型制备成功后,L组腹腔注射LPS 10 mg/kg; E组在糖尿病小鼠给予LPS处理前1 h腹腔注射EX527 5 mg/kg(溶于DMSO 0.2 ml);G组在糖尿病小鼠给予LPS处理前1 h腹腔注射银杏黄酮苷元200 mg/kg(溶于DMSO 0.2 ml);C组和D组在相同时点腹腔注射2%DMSO 0.15 ml。收集24 h尿液测定24h尿量和24 h尿蛋白浓度,眼球取血检测血清肌酐(Scr)和尿素氮(BUN)浓度。取肾组织后采用ELISA法检测IL-1β、IL-18浓度,硝酸还原酶法检测一氧化氮(NO)含量,铁离子抗氧化能力法检测总抗氧能力(T-AOC),qPCR和Western blot法检测Si...     

3D打印载PaMZ/BMP-2的nHA抗结核人工骨在体内成骨效果及药物缓释性能研究

【摘要】 目的：评估3D打印的载普托马尼（Pa）、莫西沙星（M）、吡嗪酰胺（Z）组合抗结核药物（PaMZ）和骨形态发生蛋白-2（bone morphogenetic protein-2,BMP-2）的纳米羟基磷灰石（nano-Hydroxyapatite,nHA）抗结核人工骨在体内的成骨效果及药物缓释性能。方法：取40只新西兰兔,随机分为4组,每组10只,构建脊柱骨缺损模型。其中A组植入载PaMZ/BMP-2的nHA人工骨（实验组）;B组植入空白nHA人工骨（阴性对照组）;C组植入自体髂骨（阳性对照组）;D组不植骨（空白组）。观察动物一般情况,监测体温、体重、肝肾功能;术后4、8、12周行螺旋CT三维重建观察骨缺损修复情况,并进行CT-Hedberg评分;术后8、12周行组织大体观察及植入物-骨缺损界面组织病理学观察。取SD大鼠100只,随机分为2组,实验组植入载PaMZ/BMP-2的nHA人工骨,对照组植入空白nHA人工骨,采用高效液相色谱法分别检测材料周围肌肉组织和血浆中术后不同时间点的药物浓度。结果：所有动物模型构建成功,无切口感染、死亡等情况发生。兔模型中各组动物的体温、体重及肝肾功能指标正常,组间比较均无统计学差异（P＞0.05）;CT-Hedberg评分显示在术后4、8、12周时,A组与C组相比均无统计学差异（P＞0.05）,其余各组间在不同时间点相比均存在统计学差异（P<0.05）。大体标本显示12周时A组人工骨完全被新骨包埋或替代,但B组仍有部分人工骨裸露。对植入物-骨缺损界面的骨小梁总面积定量分析显示在术后8、12周时,A组和C组相比无统计学差异（P＞0.05）,其余各组间相比均有统计学差异（P＜0.05）。SD大鼠模型对照组未测出药物成分,实验组Pa、M、Z三种药物在局部组织中均能缓释至少84d,血药浓度在植入载药人工骨后的初始时间点未测出,随后测得结果远低于局部组织药物浓度,二者具有正相关关系。结论：载PaMZ/BMP-2的nHA人工骨在体内具有良好的成骨性能,修复脊柱骨缺损的效果与自体髂骨相当,优于单纯的nHA人工骨支架;其在体内的缓释性能满意,血药浓度远低于局部组织药物浓度。     

AngiostatinK(1-3)裸DNA治疗人脑胶质瘤的实验研究

探讨ＡｎｇｉｏｓｔａｔｉｎＫ（１－３）〖ＡＫ（１－３）〗裸ＤＮＡ治疗人脑胶质可行性和作用机制。用脂质体包埋法将ＡＫ（１－３）基因的真表达载体ｐｃＤＮＡ－ＳＡＫ（１－３）注入裸以下ＳＨＧ４４人脑胶质瘤的特点。ＡＫ（１－３）裸ＤＮＡ在荷瘤裸本内获得表达,裸鼠皮下胶质瘤血管生成能力显著下降,肿瘤生长受到抑制,抑瘤率达４３％。本研究证明,ＡＫ（１－３）裸ＤＮＡ能抑制人脑质瘤生长,为非病毒载体介导的抗血管生     

螺旋CT三维成像在诊断气管、支气管疾病中作用的评价

目的 评价综合运用三维成像技术在诊断气管、支气管疾病中的作用。材料与方法 选择５３直支气管以上气道疾病患者行高档螺旋ＣＴ扫描,将容积扫描数据预处理后传至计算机工作站,分别用ＣＴ仿真支气管内镜（ＣＴＶＢ）,表面遮盖显示法（ＳＳＤ）,最大强度投影（ＭＩＰ）,最小强度投影（Ｍｉｎ ＩＰ）,多平面重构（ＭＰＲ）进行重建,分析影像学表现,４例与手术标本进行对照。结果 在轴位图像基础上,气管肿瘤在ＭＰＲ、ＣＴ